Investigating the role of HMGN2 in the self-renewal and neuronal differentiation of ECCs using the CRISPR-Cas9 knockout system

Sindi, Abdulmajeed Abdulghani A. (2017) Investigating the role of HMGN2 in the self-renewal and neuronal differentiation of ECCs using the CRISPR-Cas9 knockout system. PhD thesis, University of Glasgow.

Abstract

High mobility group N (HMGN) proteins are highly dynamic nuclear proteins that bind to nucleosomes, modulate chromatin structure and regulate transcription, replication and DNA repair. Hmgn2 mRNA is highly expressed in zones of neurogenesis in the embryonic and adult brain, but its role in these areas is unclear. In order to investigate the role of HMGN2 in stem cell self-renewal and neuronal differentiation, we used the paired nickase CRISPR-Cas9 system to specifically mutate or knockout the Hmgn2 gene in P19 embryonal carcinoma cells. Loss of HMGN2 did not affect the proliferation rate or cell cycle distribution in undifferentiated P19 cells. However, an increased rate of spontaneous neuronal differentiation was observed in HMGN2 knockout cells under conditions that normally promote self-renewal. Consistent with this, gene expression analysis revealed that several pro-neural and neuronal genes were upregulated in the HMGN2 knockout cells. Loss of HMGN2 was associated with changes in the profiles of several histone modifications at specific genomic loci. Highly efficient neuronal differentiation of P19 cells was demonstrated using an adherent cell induction protocol, and it was shown that the production of HMGN2 protein increased during the differentiation process. HMGN2 knockout cells showed greater induction of several pro-neural and neuronal-specific genes during the early stages of neuronal differentiation. These results suggest that HMGN2 plays an important role in maintaining self-renewal capacity and regulating neural induction in embryonal carcinoma cells.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Additional Information: Supported by funding from the Albaha University (Faculty of applied medical sciences) and Saudi Arabian Cultural Bureau in London
• Subjects: Q Science > QR Microbiology
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Supervisor's Name: West, Dr. Katherine
• Date of Award: 2017
• Depositing User: Mrs Marie Cairney
• Unique ID: glathesis:2017-8591
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 07 Nov 2017 11:37
• Last Modified: 22 Nov 2024 12:11
• Thesis DOI: 10.5525/gla.thesis.8591
• URI: https://theses.gla.ac.uk/id/eprint/8591